XVth International Congress on Pediatric Laboratory Medicine, Munich, Nov 27–28, 2021; Poster Presentation Abstracts

Abstract

Adenosine Deaminase 2 Deficiency (DADA2) (OMIM: 607575) is a monogenic autoinflammatory disease caused by loss of function homozygous or heterozygous mutations in ADA 2 gene (previously CECR1, Cat Eye Syndrome Chromosome Region 1. A timely diagnosis is crucial to start Anti-TNF therapies that are efficacious in controlling the disease. The confirmation of DADA2 is based on DNA sequencing and enzymatic assay. It is thus very important to have robust and reliable assays that can be rapidly utilized in specialized laboratories that can centralize samples from other centers. In this paper we show a novel enzymatic assay based on liquid chromatography-tandem mass spectrometry that allows the accurate determination of the ADA2 enzyme activity starting from very small amounts of plasma spotted on filter paper (dried plasma spot). ADA2 activity was determined in dried plasma spots (DPS) from 44 healthy donors, 18 DADA2 patients and 4 carriers. ADA2 expressed mean ± ± ± ± WHO has described malnutrition as a “ global problem ” , having an adverse effect on the survival, health performance, and progression of the population group. It is highly prevalent in developing countries among children below the age of 5 years; with severe forms occurring in 1-10% and underweight observed in 20-40%. The aim of the study was to evaluate copper and zinc levels in children with protein-energy malnutrition. Serum zinc and copper were determined in thirty (30) malnourished pre-school-age children (age, 1-5 years) and thirty (30) age-and sex-matched apparently healthy well-nourished controls to evaluate the effect of protein- energy malnutrition on serum zinc and copper. Serum zinc and copper concentrations were estimated by the Atomic Absorption Spectrophotometer. BACKGROUND-AIM Multisystem Inflammatory Syndrome in Children (MIS-C) associated with Coronavirus Disease (COVID-19) is a serious condition among paediatric patients, usually under 19 years, with previous exposure to SARS-CoV-2. Clinical manifestations consist of persistent fever along with weakness, abdominal pain, vomiting and/or diarrhea and skin rashes. Although this syndrome is rare and has variable expressivity, some children can evolve from hypotension and cardiogenic shock to multiple organ dysfunction. Laboratory abnormalities include elevation of acute phase reactants and myocardial dysfunction; similar to those observed in Kawasaki disease or toxic shock syndrome, requiring a differential diagnosis. RT-PCR respiratory viruses (SARS-CoV-2, RSV, H. in fl uenzae ), blood and urine cultures: negative. RESULTS Elevation of inflammatory markers (C-reactive protein 282 mg/L, ferritin 1103 ng/mL, IL-6 152 pg/mL, fi brinogen 667 mg/dL, PCT 6.56 ng/mL); cardiac markers (NT-proBNP 966 pg/mL); D-dimer 1120 ng/mL and renal and liver function tests (creatinine 1,04 mg/dL and GPT 44 U/L). Decrease in sodium (130 Congenital Disorders of Glycosylation (CDG) are a rapidly expanding family of rare inborn errors of metabolism. The first cases were reported 40 years ago (Jaeken et al. 1980) and today more than 130 different CDG have been reported (Jaeken 2020). MPI-CDG is characterized by a de fi ciency in mannose-6-phosphate isomerase due to autosomal recessive mutations in the MPI gene coding for the phosphomannose isomerase (Niehues et al. 1998). As a consequence, the biosynthesis of GDP-mannose and the lipid-linked oligosaccharide (LLO) precursors pools, necessary for the biosynthesis of N-glycans, are reduced leading to a defect of N-glycosylation (Ichikawa et al. 2014). MPI-CDG is mainly characterized by hepatic fi brosis, enteropathy (Pedersen and Tygstrup 1980), hyperinsulinism (de Lonlay et al. 1999), venous thrombosis, and bleeding episodes with an absence of neurological symptoms (Damen et al. 2004). MPI-CDG is the one of the only CDG where clinical manifestations are improved by oral D-mannose supplementation (Westphal et al. 2001). Symptoms usually improve with age, and normal pregnancies have already been observed in MPI-CDG patients (Helander et al. 2014; Girard et al. 2020). We studied the glycosylation of a MPI-CDG patient during pregnancy without mannose supplementation using three different methods. For the first time the glycosylation of a patient with a MPI-CDG during pregnancy without mannose supplementation is monitored using the carbohydrate deficient transferrin (CDT) assay, transferrin isoelectrofocusing We report on an 11-year-old male of Ethiopian descent who at the age of 5 days was admitted to the neonatal ICU with severe dehydration and acute renal failure that resolved following treatment. He subsequently developed uncontrolled hypertension requiring readmission and was managed on multiple antihypertensive drugs and potassium supplements, with successful blood pressure control achieved following addition of amiloride. Electrolyte measurements were performed using ion selective electrodes in an Abbott Architect ci8200 instrument). Molecular testing was undertaken in this patient. All the exons and exon-intron boundaries of the alpha- [SCNN1A (GenBank NM_001038.5)], beta- [SCNN1B (GenBank NM_000336.2)] and gamma- ENaC (epithelial sodium channel) subunits ’ [SCNN1G (GenBank NM_001039.3)] genes were ampli fi ed by PCR (New England Biolabs, Ipswich, MA, USA). Fragments were sequenced using the Nimagen, BrilliantDye ™ Terminator Cycle Sequencing Kit V3.1, BRD3-100/1000 according to the manufacturer ’ s instructions. Biochemistry test results revealed hypokalaemia, hypernatraemia and metabolic alkalosis associated with suppressed aldosterone and renin concentrations. Initial genetic testing, limited to c.1815G>A (pR563Q) of the SCNN1B gene, was negative. Further testing involved sequencing of SCNN1A and SCNN1B and revealed compound heterozygous mutations in each subunit. The c.1000G>A and c.1987A>G mutations in SCNN1A were detected, while the c7G>A and c.1325G>T were identified in SCNN1B. The timely, efficient diagnosis of inherited metabolic diseases (IMDs) via “ hyphenated ” mass spectrometry applications has become standard practice. The aim of this study was to compare a gas chromatography-mass spectrometry (GC-MS) based amino acid analysis with an automated liquid chromatography tandem mass spectrometry (LC-MS/MS) application in the diagnosis of amino acid-related IMDs. This included comparing the analytical time and diagnostic capacity of the two methods by analyzing 14 anonymizedurine samples(providedby an external proficiency BACKGROUND-AIM Heatstroke is a life-threatening injury requiring neurocritical care, characterized by central nervous system dysfunction, multiorgan failure, and extreme hyperthermia, in the setting of exposure to hot weather or extreme physical exertion. presentation. CLINICAL REPORT: A 16-year-old boy was admitted in the Paediatrics Emergency Room for malaise, tiredness and increase of his dyskinesia crisis with 1-day evolution, after spending a day at a river beach. The past history comprised psychomotor development retardation, and movement disturbance with dyskinetic crises. He had been hospitalized twice with dyskinetic crises complicated by acute kidney injury (AKI) and rhabdomyolysis. Clinical observation revealed dehydration, high fever and upper limb dyskinesia. Laboratory analysis of the patient ’ s blood revealed concentrations of leucocytes 35,4x10 9 /L, creatinine (Cr) 1.61mg/ dL, creatine kinase (CK) 4549U/L, troponin T (TnT) 32ng/L, hypernatremia and hypokalaemia. A SARS-CoV-2 RT-PCR test was negative. Being diagnosed with hypernatremic dehydration and prerenal AKI owing to a heatstroke with rhabdomyolysis, an aggressive fl uid therapy was inducted. Cystic fibrosis (CF) is an inherited disorder. Because of its varied presentation involving lungs, pancreas, and repro-ductive organs, it appears in the differential diagnosis of many pediatric diseases. Diagnosis of CF by consensus criteria includes identification of clinical features or positive family history and laboratory confirmation by identification of genetic mutations or positive sweat chloride results (two separate occasions). Over 1500 mutations in the CFTR gene make molecular diagnosis difficult. In Indians, the commonest mutation ( Δ F508) occurs in lower frequency, and also rare and novel mutations exist. Sweat chloride analysis is therefore a handy alternative tool for diagnosis. CF is not as very rare as thought and is probably under-diagnosed in Indian children. In this paper, we report our experiences with sweat chloride analysis in children of the eastern region of India using a low cost indigenous method. Briefly, involve pilocarpine iontophoresis of the local sweat production, in pre-weighed filter mins, estimation of chloride by titration with mercuric chloride. The method a sensitivity of 10 mEq/L. of > mEq/L The method effects such as in a minority of BACKGROUND-AIM Tyrosinemia type 1(Tyr-1) is an inborn error of metabolism caused by defects in tyrosine metabolism and characterized by accumulation of tyrosine and the toxic metabolite, succinylacetone (SA). Treatment of Tyr-1 patients is based on niti-sinone (NTBC) administration and tyrosine- and phenylalanine-restricted diet. To prevent liver complications such as hepatocarcinogenesis, permanent monitoring of NTBC concentration in blood, SA levels, alpha-fetoprotein ( α FP) and liver function biomarkers are recommended, to be measured in each clinical control, by the current consensus guidelines. Also, the routine monitoring of these parameters allows to improve the NTBC dosage in the patients. The aim of present study was to evaluate the optimal therapeutic range of NTBC in Tyr-1 Patients. concentration in DBS was established within 14.5- 24.9 μ mol/L. Also, we explored a possible correlation of NTBC range levels with nutritional follow-up parameters (tyrosine and phenylalanine) and liver biomarkers, showing that for α FP, NTBC concentrations trends to be negatively associated. Our observational study